CAN SOME ONE EXPLAIN WHAT IS HAPPENING IN THESE TWO REACTIONS.....& I USED SAMPLES OF DNA DILUTION, THE SENSITIVITY BETWEEN THESE TWO REACTION IS ATLEAST ONE LOG.THE FIRST REACTION GAVE ME A SENSITIVITY OF 10^6 & THE SECOND ONE CAN COME UP TO ONLY 10^4

CAN SOME ONE EXPLAIN WHAT IS HAPPENING IN THESE TWO REACTIONS.....& I USED SAMPLES OF DNA DILUTION, THE SENSITIVITY BETWEEN THESE TWO REACTION IS ATLEAST ONE LOG.THE FIRST REACTION GAVE ME A SENSITIVITY OF 10^6 & THE SECOND ONE CAN COME UP TO ONLY 10^4

PLEASE THROW SOME DISCUSSION ON THIS.

FIND PICS.

Without knowing the sequences of the primers or whether there is mispriming evident on your gels, I'm guessing that a) there's mispriming at the lower temperature or there are primer dimers forming at the lower temperature. Either way, it looks like you might as well forget the touchdown part and just do 35 cycles at the higher temperature.